离子交换树脂提取纯化杆菌肽的工艺优化

Optimization in Extraction and Purification of Bacitracin by Ionic Exchange Resin

首先比较了732、HZ016、D152、HD-2、D061、D001-CC、D296、D301等8种不同类型的离子交换树脂对发酵液中杆菌肽的吸附和解吸效果,选出适合提取纯化杆菌肽的树脂;然后以交换容量和洗脱收率为指标,对交换pH值、交换流速和洗脱条件进行优化。结果表明,大孔强酸型阳离子交换树脂D061对杆菌肽的纯化效果较好;最佳工艺条件为:交换pH值5.0~6.0,交换流速2.5BV·h^(-1),先用0.5%氨水以流速0.5BV·h^(-1)洗脱2倍树脂体积,再用3.5%氨水洗脱3倍树脂体积,在此条件下,洗脱液中杆菌肽A含量可达66.2%,杆菌肽洗脱收率可达91%以上。所得洗脱液串联凝胶型001X4阳离子交换树脂柱进行脱盐处理,再经过脱色、萃取、结晶、干燥等精制步骤可得到生物效价高于74U·mg^(-1)的医药级杆菌肽原料药。该工艺简单可行,收率稳定,纯化效率高,为工业化生产杆菌肽奠定了理论基础。

We firstly compared the adsorption and desorption efficiencies of eight kinds of ionic exchange resins, namely 732, HZ016, D152, HD- 2, DO 61, D001- CC, D296, and D301 on bacitracin in fermentation broth, and selected the suitable resin for extraction and purification of bacitracin. Then,we optimized exchange pH val- ue,exchange flow rate,and elution conditions by using exchange capacity and elution yield as indexes. Results showed that macroporous strong acid cation exchange resin D061 was the best one to extract and purify bacitra- cin. The optimal process conditions were as follows：exchange pH value was 5.0-6.0,exchange flow rate was 2.5 BV · h^-1 ,eluting 2 times of the resin volume with 0.5% ammonia with flow rate of 0.5 BV · h-1 firstly, then eluting 3 times of the resin volume with 3.5 % ammonia. Under above conditions, the content of bacitracin A in eluent reached 66.2 %,and elution yield reached over 91%. After desalination by gel type cation exchange resin 001X4,and following purification steps such as decolorization, extraction, crystallization, and drying, we obtained pharmaceutical grade bacitracin with the titer of 74 U · mg^-1. The process is easy and feasible with stable yield and high purification efficiency,which lays a theoretical foundation for industrial production of bacitracin.